Abstract
Conclusion:
These results show for the first time that the RAS/RAF/ERK1/2 MAPK signalling pathway is active and involved in p21-mediated cell cycle arrest in human cholesteatoma epithelium.
Objective:
In a previous report we have demonstrated that the epithelium in human cholesteatoma is characterized by high p53-dependent p21 expression. The RAS/RAF/extracellular signal-regulated kinase (ERK)1/2 mitogen-activated protein kinase (MAPK) signalling pathway can induce p21 expression and subsequent cell cycle arrest via p53-dependent or -independent mechanisms. We designed the present study to investigate whether the RAS/RAF/ERK1/2 MAPK signalling pathway is involved in p53-dependent and p21-mediated cell cycle arrest in human cholesteatoma.
Material and methods:
A total of 18 cholesteatoma samples and 18 paired control retro-auricular skin samples were immunohistochemically stained for p53, p21, phosphorylated ERK1/2 (pERK1/2) and total ERK1/2. Positive cells were counted by means of digital image analysis. Double-label fluorescence immunohistochemistry was performed to demonstrate co-expression of p21 and pERK1/2.
Results:
Protein expression of p53, p21 and pERK1/2 differed significantly between cholesteatoma epithelium and retro-auricular skin (p <0.01). In cholesteatoma, co-expression of p21 and pERK1/2 was prominent, whereas in retro-auricular skin there was hardly any co-expression. Positive correlations were found between p53 and p21 (p =0.003) and between p21 and pERK1/2 (p =0.013).
MeSH terms
-
Antibodies, Monoclonal / immunology
-
Cell Cycle / genetics
-
Cell Cycle / physiology*
-
Cell Cycle Proteins / genetics
-
Cell Cycle Proteins / immunology
-
Cell Cycle Proteins / metabolism*
-
Cholesteatoma, Middle Ear / genetics
-
Cholesteatoma, Middle Ear / metabolism*
-
Cholesteatoma, Middle Ear / pathology
-
Cyclin-Dependent Kinase Inhibitor p21
-
Epithelium / metabolism
-
Epithelium / pathology
-
Extracellular Signal-Regulated MAP Kinases / genetics
-
Extracellular Signal-Regulated MAP Kinases / immunology
-
Extracellular Signal-Regulated MAP Kinases / metabolism
-
Genes, cdc / physiology
-
Humans
-
Immunohistochemistry
-
MAP Kinase Kinase Kinases / genetics
-
MAP Kinase Kinase Kinases / immunology
-
MAP Kinase Kinase Kinases / metabolism
-
Membrane Glycoproteins / metabolism
-
Mitogen-Activated Protein Kinase 1 / immunology
-
Mitogen-Activated Protein Kinase 1 / metabolism
-
Mitogen-Activated Protein Kinase 3 / immunology
-
Mitogen-Activated Protein Kinase 3 / metabolism
-
Mitogen-Activated Protein Kinases / immunology
-
Mitogen-Activated Protein Kinases / metabolism*
-
Phosphorylation
-
Signal Transduction / genetics*
-
Tumor Suppressor Protein p53 / genetics
-
Tumor Suppressor Protein p53 / immunology
-
raf Kinases / genetics
-
raf Kinases / metabolism
Substances
-
Antibodies, Monoclonal
-
CDKN1A protein, human
-
Cell Cycle Proteins
-
Cyclin-Dependent Kinase Inhibitor p21
-
Membrane Glycoproteins
-
Tumor Suppressor Protein p53
-
raf Kinases
-
Extracellular Signal-Regulated MAP Kinases
-
Mitogen-Activated Protein Kinase 1
-
Mitogen-Activated Protein Kinase 3
-
Mitogen-Activated Protein Kinases
-
MAP Kinase Kinase Kinases
-
Ras-dependent mitogen-activated protein kinase kinase kinase